The overall objective of this application is to develop a shared resource at the Medical College of Wisconsin for the analysis of biomolecular structure and dynamics using high-resolution NMR spectroscopy. Funds are requested for a 500 MHz NMR spectrometer, which will be made available to an expanding base of users in the biomedical research community of southeastern Wisconsin. This instrument will support and extend existing NIH projects for 4 major users and 12 minor users from 8 institutions. The major projects are funded by 6 NIH grants, and minor users are supported by various agencies. The NMR needs of this user group exceed the time available on existing 600 MHz instruments at the Medical College of Wisconsin. A key requirement for investigations of protein dynamics is relaxation data acquired at two different magnetic field strengths, and the cryoprobe-equipped 500 MHz instrument requested in this application will provide an optimal solution that is not readily available at other nearby NMR facilities. Major projects include: (1) Dynamics of the lymphotactin native state structural equilibrium (Volkman), (2) Structural basis of selective activation of the anthrax-killing enzyme PlyG (Volkman), (3) Target screening and optimization for the Center for Eukaryotic Structural Genomics (Volkman), (4) Structural analysis of mannose-6-phosphate receptors (Dahms), (5) Bicarbonate enhances peroxidation of SOD/ALS mutants (Kalyanaraman) and (6) Solution structure of the PECAM-1 cytoplasmic domain (Newman). The 500 MHz instrument will be the first component of a major expansion to the MCW Biomolecular NMR Center, in conjunction with plans for a new Center for Structural Biology. Accordingly, MCW has committed $250,000 in matching funds for the instrument and $500,000 for construction of the NMR facility. Relevance: Alterations in protein folding, structure and dynamics form the basis of many human diseases. Projects using the requested NMR instrument will answer questions about inherited mutations that cause ALS (Lou Gehrig's disease) or light-chain amyloidosis (AL), show how engineered proteins can be used to treat lysosomal storage diseases (Gaucher, Fabry, and Tay-Sachs, for example), and define the structure of a selective anthrax-killing enzyme. Other studies will exploit differences in magnetic field strength to measure fluctuations in three-dimensional protein structures that control cellular signals in cancer and the immune response. PUBLIC HEALTH RELEVANCE: Alterations in protein folding, structure and dynamics form the basis of many human diseases. Projects using the requested NMR instrument will answer questions about inherited mutations that cause ALS (Lou Gehrig's disease) or light-chain amyloidosis (AL), show how engineered proteins can be used to treat lysosomal storage diseases (Gaucher, Fabry, and Tay-Sachs, for example), and define the structure of a selective anthrax-killing enzyme. Other studies will exploit differences in magnetic field strength to measure fluctuations in three-dimensional protein structures that control cellular signals in cancer and the immune response.